Department of Agricultural Chemistry, Sunchon National University, Suncheon 540-742, South Korea.
Appl Biochem Biotechnol. 2012 Sep;168(2):375-82. doi: 10.1007/s12010-012-9781-x. Epub 2012 Jun 23.
XynX of Thermoanaerobacterium sp. [corrected] is a large, multimodular xylanase of 116 kDa. An Escherichia coli transformant carrying the entire xynX produced three active truncated xylanase species of 105, 85, and 64 kDa intracellularly. The Bacillus subtilis WB700 transformant with the xynX, a strain deficient in seven proteases including Vpr, secreted two active truncated xylanase species of 65 and 44 kDa. The B. subtilis WB800 transformant with xynX, a strain deficient in eight proteases including Vpr and WprA, secreted more active enzymes, 8.46 U ml(-1), mostly in the form of 105 and 85 kDa, than the WB700 transformant, 6.93 U ml(-1). This indicates that the additional deletion of wprA enabled the WB800 to secrete XynX in its intact form. B. subtilis WB800 produced more total enzyme activity than E. coli (1,692 ± 274 U vs. 141.9 ± 27.1 U), and, more importantly, secreted almost all the enzyme activity. The results suggest the potential use of B. subtilis WB800 as a host system for the production of large multimodular proteins.
热厌氧菌[已更正]的 XynX 是一种分子量为 116 kDa 的大型多结构域木聚糖酶。携带完整 xynX 的大肠杆菌转化体在细胞内产生了三种活性截断的木聚糖酶,分子量分别为 105、85 和 64 kDa。带有 xynX 的枯草芽孢杆菌 WB700 转化体,该菌株缺乏包括 Vpr 在内的七种蛋白酶,分泌了两种活性截断的木聚糖酶,分子量分别为 65 和 44 kDa。带有 xynX 的枯草芽孢杆菌 WB800 转化体,该菌株缺乏包括 Vpr 和 WprA 在内的八种蛋白酶,比 WB700 转化体分泌更多的活性酶,8.46 U ml(-1),主要以 105 和 85 kDa 的形式存在,比 WB700 转化体(6.93 U ml(-1))更多。这表明 wprA 的额外缺失使 WB800 能够以完整形式分泌 XynX。枯草芽孢杆菌 WB800 产生的总酶活性比大肠杆菌(1692 ± 274 U 对 141.9 ± 27.1 U)更多,更重要的是,几乎分泌了所有的酶活性。结果表明,枯草芽孢杆菌 WB800 可能作为一种生产大型多结构域蛋白的宿主系统。